Wahba Amir E, Fedele Denise, Gebril Hoda, AlHarfoush Enmar, Toti Kiran S, Jacobson Kenneth A, Boison Detlev
Department of Neurosurgery, Robert Wood Johnson Medical School, Rutgers University, Piscataway, New Jersey 08854, United States.
Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 8A, Room B1A-19, Bethesda, Maryland 20892-0810, United States.
ACS Pharmacol Transl Sci. 2021 Feb 16;4(2):680-686. doi: 10.1021/acsptsci.1c00008. eCollection 2021 Apr 9.
DNA methylation has a major role in cancer, and its inhibitors are used therapeutically. DNA methylation depends on methyl group flux through the transmethylation pathway, which forms adenosine. We hypothesized that an adenosine kinase isoform with nuclear expression (ADK-L) determines global DNA methylation in cancer cells. We quantified ADK-L expression (Western Blot) and global DNA methylation as percent 5-methyldeoxycytidine (5mdC, LC-MS/MS) in three cancer lines (HeLa, HepG2, and U373). ADK-L expression and global DNA methylation correlated positively with the highest levels in HeLa cells compared to U373 and HepG2 cells. To determine whether ADK increases global DNA methylation and to validate its potential therapeutics, we treated HeLa cells with potent ADK inhibitors MRS4203 and MRS4380 (IC 88 and 140 nM, respectively). Both nucleosides, but not a structurally related poor ADK inhibitor, significantly reduced global DNA methylation in HeLa cells in a concentration-dependent manner. Thus, ADK-L is a potential target for the therapeutic manipulation of DNA methylation levels in cancer.
DNA甲基化在癌症中起主要作用，其抑制剂被用于治疗。DNA甲基化依赖于通过转甲基途径的甲基通量，该途径形成腺苷。我们假设一种具有核表达的腺苷激酶同工型（ADK-L）决定癌细胞中的整体DNA甲基化。我们在三种癌细胞系（HeLa、HepG2和U373）中通过蛋白质免疫印迹法定量ADK-L表达，并通过液相色谱-串联质谱法将整体DNA甲基化定量为5-甲基脱氧胞苷（5mdC）的百分比。与U373和HepG2细胞相比，HeLa细胞中ADK-L表达和整体DNA甲基化呈正相关且水平最高。为了确定ADK是否增加整体DNA甲基化并验证其潜在治疗作用，我们用强效ADK抑制剂MRS4203和MRS4380（分别为IC 88和140 nM）处理HeLa细胞。这两种核苷，而非结构相关的低效ADK抑制剂，以浓度依赖的方式显著降低了HeLa细胞中的整体DNA甲基化。因此，ADK-L是癌症中DNA甲基化水平治疗调控的潜在靶点。
